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FIGURE 4. The ribavirin-mediated regulation of CHS responses and expression of the costimulatory molecules, B7-1 and B7-2, in BALB/c (left panels) and C57BL/6 (right panels) mice is dependent on IL-10. A, CHS responses were induced in five groups of five mice of either strain by sensitization (day 0) and challenge (day 6) with DNFB (Primed), as described in Materials and Methods. Twenty-four hours before challenge, two Primed BALB/c groups and one Primed C57BL/6 group were administered IL-10 (1 mg) i.p., and one Primed C57BL/6 group was administered anti-IL-10 Ab (50 mg) i.p. At the time of challenge with 0.12% DNFB, two BALB/c groups, one Primed group (Primed 1 RIB), and one Primed group injected with IL-10 (Primed 1 RIB 1 IL-10) were administered 5 mg of ribavirin (RIB) in PBS by i. p. injection. Of the C57BL/6 mice, one Primed group (Primed 1 RIB) and one Primed group injected with anti-IL-10 Ab (Primed 1 RIB 1 IL-10 Ab) were administered 5 mg of ribavirin. All mouse groups, including the remaining DNFB-primed, IL-10 group from each strain (Primed 1 IL-10), were challenged on the ears with 0.12% DNFB on day 6. The ear swelling in all groups (including an unsensitized control group (Naive)) was measured after 24 h. The data shown as mean ear swelling (mm 3 1022) 6 SD are representative of two experiments with five to six animals in each test group. p, p 5 0.0002 for group comparisons against DNFB-primed mice alone. pp, p 5 0.0003 for comparison against ribavirin-treated DNFB-primed mice. B, B7-1 and B7-2 mRNA expression, 24 h post-DNFB challenge, was determined in total RNA isolated from the LNC of individual DNFB-primed mice (DNFB) and DNFBprimed mice treated with either IL-10 (DNFB 1 IL-10), ribavirin (DNFB 1 RIB), or ribavirin and IL-10 (BALB/c) (DNFB 1 IL-10 1 RIB) or ribavirin and anti-IL-10 Ab (C57BL/6) (DNFB 1 anti-IL-10 Ab 1 RIB). B7-1 and and B7-2 mRNA levels were assessed following RT-PCR and Southern blot analysis with specific 32P-labeled DNA probes. RNA samples from an unprimed mouse challenged with DNFB (Naive) and a DNFB-primed C57BL/6 mouse injected with isotype control Ab (Ig) were also included. Autoradiography data are representative of two separate experiments using three mice per treatment group. C, The relative changes in B7-1 (hatched bars), and B7-2 (solid bars) mRNA expression from the mouse groups in B are presented as densitometric readings. Primed, Primed 1 RIB, Primed 1 IL-10, and Primed 1 RIB 1 IL-10 (BALB/c) and Primed 1 RIB 1 anti-IL-10 Ab (C57BL/6) mouse groups are normalized